6-styrylpyrazolo(3,4-d)pyrimidinones and pyrimidines

ABSTRACT

1. A COMPOUND OF THE FORMULA   1-R1,3-R2,4-(O=),6-(R3-PHENYLENE-CH=CH-)-PYRAZOLO(3,4-D)-   PYRIMIDINE   WHEREIN R1 IS LOWER ALKYL, CYCLO-LOWER ALKYL OR PHENYLLOWER ALKYL; R2 IS HYDROGEN OR LOWER ALKYL AND R3 IS HYDROGEN, LOWER ALKYL, HALOGEN OR TRIFLUOROMETHYL.

United States Patent 3,847,908 G-STYRYLPYRAZOLO[3,4-d]PYRIMIDINONES AND PYRIMIDINES Hermann Breuer and Uwe D. Treuner, Regensburg, Germany, assignors to E. R. Squibb & Sons, Inc., Princeton, NJ. No Drawing. Filed Mar. 5, 1973, Ser. No. 337,807 Int. Cl. C09d 23/14 US. Cl. 260-240 D 13 Claims ABSTRACT OF THE DISCLOSURE 6 Styrylpyrazolo[3,4-d]pyrimidin-4-ones of the general formula and the 6-styrylpyrazolo[3,4-dJpyrimidines which are derived from them, having the general formula are useful as antimicrobial agents and also have antiinfiammatory and membrane stabilizing properties.

SUMMARY OF THE INVENTION This invention relates to fi-styrylpyrazolo[3,4-d1pyrimidin-4-ones which have the formula and to the 6-styrylpyrazolo[3,4-d1pyrimidines which are obtained from them and which have the formula (II) R 3,847,908 Patented Nov. 12, 1974 ice together is morpholino, piperidino or lower alkylpiperidino.

DETAILED DESCRIPTION OF THE INVENTION The lower alkyl groups in the various radicals represented by the symbols are straight or branched chain aliphatic hydrocarbon groups having one to seven carbon atoms, for example, methyl, ethyl, propyl, isopropyl, butyl, t-butyl, amyl and the like, the one to four carbon groups being preferred.

The cyclo-lower alkyl groups are the four to six carbon alicyclics cyclobutyl, cyclopentyl and cyclohexyl, the last two being preferred.

The phenyl-lower alkyl groups include lower alkyl groups of the kind described above, but benzyl and phenethyl are preferred, especially the first.

All four of the common halogens are contemplated. Chlorine, bromine and fluorine are preferred within that group, but especially chlorine.

The basic radical includes such groups as amino, lower alkylamino, e.g., methylamino, ethylarnino, propylamino, butylamino, etc., di-lower alkylamino, e.g., dimethylamino, diethylamino, dipropylamino, dibutylamino, etc., hydroxy-lower alkylamino, e.g., hydroxyethylamino, di(hyclroxy-lower alkyl) amino, e.g., di(hydroxyethyl)amino, di(lower alkyl) amino-lower alkyl, e.g., dimethylaminomethyl, diethylaminomethyl, dipropylaminomethyl, dimethylaminoethyl, diethylaminoethyl, dipropylaminoethyl, dimethylaminopropyl, and the like (the one to three carbon al kyls are preferred, especially those groups named). Preferably there is only one of the bulkier groups. In addition, R and R join together with the nitrogen to form one of the heterocyclic groups morpholino, piperidino or lower lalkylpiperidino, e.g., N -rnethylpiperidino or N -ethyl piperidino.

When there is a basic side chain, in the 4-position, acid addition salts are formed and these are also within the scope of the invention.

The lower alkyl groups represented by the various symbols are saturated hydrocarbon radicals of up to seven carbon atoms like methyl, ethyl, propyl, isopropyl, butyl, isobutyl, t-butyl and the like. The cyclo-lower alkyl groups are three to seven alicyclics cyclopropyl to cycloheptyl. The five and six carbon members are preferred. All four halogens are included but chlorine and bromine are preferred.

DETAILED DESCRIPTION OF THE INVENTION is produced by reacting a dicyano compound of the formula (IV) R2 with a hydrazide H NNH-R in alcohol at about reflux temperature.

This 5-amino-4-cyanopyrazole is made to react with a cinnamoyl halide of the formula x ti-on=crr wherein X is halogen, preferably chlorine, in an inert organic solvent such as dioxane, in the presence of an organic base such as pyridine, to obtain a S-cinnamoylamino-4-cyanopyrazole of the formula (VII) The product of formula II is then obtained by treating the halogenated compound of formula VII with an amine (V III) e.g., in an inert organic solvent like toluene, preferably at an elevated temperature, e.g., up to reflux temperature.

The products with basic side chains, especially when there is a di-lower alkylamino-lower alkyl group, form the acid addition salts referred to above, for example, by reaction with an ethereal solution containing an equivalent proportion of the appropriate acid. The acid addition salts include the commonly known salts such as hydrohalides, especially hydrochloride and hydrobromide, sulfate, nitrate, phosphate, and the like as well as organic acid salts like acetate, citrate, succinate, benzoate, salicylate, malate, citrate and the other well known members.

Certain of the compounds herein exist in isomeric forms and these are included within the scope of the invention.

The new compounds of this invention are useful as antimicrobial agents and may be used to combat infections in animal species, such as mice, rats, dogs, guinea pigs and the like, due to organisms such as Trichomonas vaginalis, Candida albicans, T riclzomonas foetus, Staphylococcus aureus, Salmonella sclzomnuelleri, Proteus vulgaris, Escherichia coli or T richophyton mentagrophytes. For example, a compound of formula I or of formula II or physiologically acceptable acid addition salt thereof may be administered orally to an infected animal, e.g., to a mouse, in an amount of about to 25 mg. per kg. per day in 2 to 4 divided doses. These may be conventionally formulated in a tablet, capsule or elixir containing about to 250 mg. per dosage unit, by compounding the active substance or substances with the conventional excipient, vehicle, binder, perservative, flavor, etc. as called for by accepted pharmaceutical practice. They may also be applied topically, e.g., to dermatophytosis in a guinea pig,

in a lotion, salve or cream at a concentration of about 0.01 to 3 percent by weight.

The new compounds of this invention, especially those of formula II, also have antiinflammatory properties and are useful as antiinflammatory agents, for example, to reduce local inflammatory conditions such as those of an edematous nature or resulting from proliferation of connective tissue in various mammalian species such as rats, dogs and the like when given orally in dosages of about 5 to mg./kg./day, preferably 5 to 25 mg./kg./day, in single or 2 to 4 divided doses, as indicated by the carageenan edema assay in rats. The active substance may be utilized in compositions such as tablets, capsules, solutions or suspensions containing up to about 300 mg. per units of dosage of a compound or mixture of compounds of formula II or physiologically acceptable acid addition salt thereof. They may be compounded in conventional manner with a physiologically acceptable vehicle or carrier, excipient, binder, preservative, stabilizer, flavor, etc. as called for by accepted pharmaceutical practice. Topical preparations containing about 0.01 to 3 percent by weight of active substance in a lotion or cream may also be used. The compounds also have membrane stabilizing properties.

The following examples are illustrative of the invention. Additional members of the class are produced in the same manner by simple substitution of the reactants having the appropriate R group. All temperatures are on the centigrade scale.

Example 1 (a) 1-Methyl-4-cyano-5-aminopyrazole.-To a solution of grams of 98% methyl hydrazine and 700 ml. of ethanol are added 127 gms. of ethoxy methylene malonitrile in such a manner that the solution remains at the boiling point. After completion of the addition the reaction mixture is heated to reflux for 30 minutes. The product is permitted to crystallize overnight in the refrigerator. After filtering under suction and washing with a small amount of cold ethanol 103 gms. of crude 5-amino-4-cyano-l-methylpyrazole remains as yellow crystals, mp. 216-218".

(b) Trans-l-methyl 4 cyano-S-cinnamoylaminopyrazole.6.l g. of 1-methyl-4-cyano-5-aminopyrazole in 250 ml. of dioxane and 2.5 ml. of pyridine are stirred for one hour with 8.6 g. of cinnamoyl chloride at 10-15 and then heated at for an additional 30 minutes. After distilling off the solvent, the oily residue is treated with methanol-water (1:1) until it solidifies. The light yellow solid is then filtered under suction, thoroughly washed with water, dried and recrystallized from methanol; yield 7 g. of light yellow crystals of trans-1-methyl-4- cyano-5-cinnamoylaminopyrazole, mp. 161-165 The following additional 1-methyl-4-cyano-S-cinnamoylaminopyrazoles are obtained by the foregoing procedure by substituting for the cinnamoyl chloride the chloro substituted cinnamoyl chloride and in the case of compounds (c) to (f) which follow, substituting 1,3-dimethyl-4-cyano- S-aminopyrazole for the 1 methyl 4 cyano-S-aminopyrazole:

R2 CN ll N N t @H-CH C? M.P.. R R3 degrees Appearance CH H 222-225 Yellow crystals. CH o-Cl 240-242 D0. CH: m-Cl 188-190 D0. CH; p-Cl 214-217 Do. H o-Cl 217-219 D0. H m-Cl -172 Do. H p-Cl 194498 D0.

(j) 1,5-Dihydro-1-methyl 6 styryl 7 4H pyrazolo [3,4-d]pyrimidin-4-one.6.2 g, of 1-methyl-4-cyano-5- cinnamoylaminopyrazole, 90 ml. of 3% hydrogen peroxide and 2.5 g. of potassium hydroxide are maintained at 70-75 for three hours. After cooling, the reaction mix- 6 methyl-6-(m-chlorostyryl)-4I-I pyrazolo[3,4 d]pyrimidin-4-one in 30 ml. of phosphorus oxychloride and 30 g. of phosphorus pentachloride are maintained at reflux temperature for three hours. After working up the product as in part (a) 17.2 g. of white crystalline 4-chloro 6-(mture is acidified with acetic acid and the 1,5-dihydro-l- 5 chlorostyryl)-l methyl-lH pyrazolo[3,4 d]pyrimidine methyl-6-styryl-4H-pyrazolo[3,4 dJpyIimidin 4 one are obtained, mp. 172-l74. precipitates as a light yellow precipitate. Recrystallization The following additional compounds are obtained by from methyl glycol gives 4 g. of light yellow needles, mp. the same procedure from the products of Example 1: 279-282. C1

By utilizing the intermediate (e) to (i), respectively in I the procedure of part (j) above, the following additional N compounds are obtained. I s

N J 0 CH=CH II 1r W N CH=CH' MP" f N/ R: R: degrees Appearance R1 CH3 o-Cl 152-154 White powder.

CH3 m-Cl 140-142 Yellow needles. M P CH5 p-Cl 128-130 White crystals. R2 R3 degrees Appearance (f) 4-(n-Butylamino)-6-(m-ehlorostyryl) 1 methylgg: E 5 82 g; crystalslH-pyrazolo[3,4-d]pyrimidine.3 g. of 4-chl oro 6-(m- CH3 m-Cl 308-311 Do. chlorostyryl)1-methyl-1H-pyrazolo[3,4 d]pyr1m1d1ne 1n E 5& 23:33? ml. of toluene and 1.45 g. of n-butylamine are refluxed H m-Cl 297-298 Do. for one hour. The reaction mixture is filtered and the H 307-310 Whlte mwde" filtrate is concentrated. The solid yellow residue is crystal- 1 2 3O lized from methanol to obtain 1.9 g. of 4-(n-butylamino)- Examp e 6-(m-chlorostyryl)l methyl 1H pyrazolo [3,4 d] (a) 4-Chl0r0-6 -(p chlorostyryl) 1 methyl lH- pyrimidine as light yellow needles, mp. 162164. pyrazolo[3,4-d]pyrimidine.3.2 g. of 1,5 dihydro l- By utilizing the products of Examples 1 and substituting methyl-6-(p -chlorostyryl)-4H-pyrazolo[3,4 d]pyn'midinfor the n-buytlamine the appropriate amine in the fore- 4-one in 30 ml. of phosphorus oxychloride is refluxed with going procedure, the following additional products are 3 g. of phosphorus pentachloride for three hours. The exobtained:

RI i R R5 1| L N R3 \N NJ CH:CH*@

M.P., R1 R: R3 R5 degrees Appearance (g)... CH3 H p-Ol -NH-(CH2)aCH3 69-72 Light yellow crystals.

(h) CH3 CH3 o-Cl N/ w) 114-115 White powder.

(i) CH3 CH3 m-Cl -NH(CH2)3CH3 130432 White crystals. 3 CH3 H p-Cl -NH(CH2)3)2 88-90 Do. (k) CH3 CH3 0-01 --NH-(CH2)aN(OHa)2 128430 Do. 1) CH3 H o-Cl -NH-(CHz)3N(CHa)z 90 Do. (m)... CH3 H 0-01 NH(CH2)aCHa -122 D0. (n).-- CH3 CH3 0-01 -NH(OH2)aCHa 4 123 Do.

cess phosphorus oxychloride is then distilled off and the The hydrochloride salts of the products of (k) and (l) cooled residue is stirred with ice water whereupon White above are formed by dissolving the crystals in ethanol, crystals separate. After thorough washing with ice Water, adding an equivalent amount of hydrogen chloride in the crystals are dried and recrystallized from cyclohexane. ether solution, then precipitating the salt bythe addition 2.8 g. of White crystalline 4-chloro-6-(p-chlorostyryl)-1- of more ether. methyl-lH-pyrazolo[3,4-d1pyrimidine are obtained, m.p. Example 3 152-154".

(b) 4-chloro-6-(m chlorostyryl) 1 methyl 1H- 75 The following additional products are obtained by the pyrazolo[3,4-d]pyrimidine.20 g. of 1,5 dihydro 1- procedure ofExample 1 by utilizing the R R substituted-4-cyano-S-aminopyrazole and R -substituted cin- What is claimed is: namoyl chloride: 1. A compound of the formula (b). H o-Cl wherein R is lower alkyl, cyclo-lower alkyl or phenyl- S lower alkyl; R is hydrogen or lower alkyl and R is (c) CH: H hydrogen, lower alkyl, halogen or trifiuoromethyl.

@- 15 2. A compound of the formula ((1)----- OH: H R

CH2CI'I3 l Example 4 The following additional products are obtained by the procedure of Example 2 from the products of Example 1 and Example 3 but substituting for the n-butylamine the wherein and R3 have the Same meanmg as m Claim 1 and R and R each is hydrogen, lower amme alkyl, hydroxy-lower, di-lower alkylamino-lower alkyl or together with the nitrogen are morpholino, piperi- HN 30 dino or lower alkylpiperidino,

and acid addition salts of the basic members.

H N m N All 3 R1 R3 R5 H p-Cl --N(C1H5OH)2 H p-Cl -NH-(CH2)2-N(C:H5)3 OH; H NH:

(d) CH; H In-Cl N/ \N (e) CH3 H m CFa N11,

r OH H -c1 O a m N \N-CH;;

(g) CH; CH: o-Cl (h) CH H NH(CH2)2OH i H H (j) S H 0-01 N(C2H5)1 (k) 3 CH: H -NH-(CH2)aCHa (l) CzHs CzHs p-Cl -NH-(CH2)3-N(CH3)I (In) CH: CH: O-CH:

N N-CH:

(n) CH CH1 o-Br ---N(CH:):

(0) CH: CH: p-Cl CHCHOH 3. A compound as in Claim 1 wherein R and R each is lower alkyl and R is halogen.

4. A compound as in Claim 1 wherein R is lower alkyl, R is hydrogen and R is halogen.

5. A compound as in Claim 3 wherein each lower alkyl group is methyl and the halogen is chlorine.

6. A compound as in Claim 2 wherein R and R each is lower alkyl, R is halogen and is lower alkylamino.

7. A compound as in Claim 2 wherein R is lower alkyl, R is hydrogen, R is halogen and is di-lower alkylamino-lower alkyl.

11. A compound as in Claim 2 wherein R is lower alkyl, R is hydrogen, R is halogen and is di-lower alkylamino-lower alkyl.

12. A compound as in Claim 10 wherein each lower alkyl group is methyl, the halogen is chlorine and the dilower alkylamino-lower alkyl group is dimethylaminopropylamino.

13. A compound as in Claim 11 wherein the lower alkyl group is methyl, the halogen is chlorine and the dilower alkylamino-lower alkyl group is dimethylaminopropylamino.

References Cited UNITED STATES PATENTS 3,165,520 1/1965 Schmidt et al. 260256.4 F 3,211,732 10/1965 Schmidt et al. 260256.4 F 3,732,225 5/1973 Breur et al. 260-256.4 F

JOHN D. RANDOLPH, Primary Examiner US. Cl. X.R.

UNITED STATES PATENT AND TRADEMARK OFFICE CERTIFICATE OF CORRECTION PATENT NO. 3,847,908

DATED November 12, 1974 INVENTOR(S) Hermann Breuer, Uwe D. Treuner It is certified that error appears in the above-identified patent and that said Letters Patent are hereby corrected as shown below:

Column 2, line 1, (2 occurrences) line 39, line 40,

(2 occurrences) line 41 piperidino should be "piperazino". Column 4, line 34 malo should be "malono". Column 6, line 5 17.2 g. should be "17.3 g. Column 6, in the second table (j) under 4 n N\R (cH 2 should be NH (CH -N(CH 2 5 R Column 7, Example 4 (e) under N 4 NH should be IINHZII 9 Column 8, line 30 (2 occurrences) --piperidino-- should be "piperazino".

. Signed and Scaled this Thirty-first Day of August 1976 A nest:

RUTH C. MASON C. MARSHALL DANN Arresting Officer Commissioner uj'Parems and Trademarks 

1. A COMPOUND OF THE FORMULA 